Serogroup identification and Virulence gene characterization of Listeria monocytogenes isolated from chicken carcasses

Document Type : Research Articles

Authors

1 Ferdowsi university of mashhad

2 ferdowsi university of mashhad

Abstract

Listeria monocytogenes is an important food borne pathogen with a high case fatality rate. Among 13 different serotypes of this bacterium, 4 serotypes (1/2a, 1/2b, 1/2c, and 4b) were considered as the main cause of listeriosis outbreaks. The aim of this study is to identify the major serotypes and virulence genes in Listeria monocytegenes isolated from chicken carcasses which were collected from different supermarkets and butcheries in Mashhad. Among the 80 isolated Listeria spp., most of them were identified as L. monocytogenes (36 out of 80). Most of the L. monocytogenes isolates belonged to serogroup llb (52.77%) which contains 1/2b and 3b serotypes. The second and third major serogroups were IVa (27.77%) and IIa (16.66%). Serogroup IVb (2.77%) which contains 4b serotype was the fourth major isolate. In order to differentiate serotype 1/2a and 3a from 1/2c, amplification of flaA gene was used. L. monocytogenesis isolates were also examined for the presence of inlC, inlJ and hlyA virulence genes. 26 out of 36 isolates were positive for inlC, and inlJ, whereas hlyA gene was detected in 32 isolates. Chicken carcasses may act as a source of infectious listeriosis for humans living in this area.

Keywords


Amagliani, G., Giammarini, C., Omiccioli, E., Brandi, G., Magnani, M. (2007). Detection of Listeria monocytogenes using a commercial PCR kit and different DNA extraction methods. Food Control 18, 1137–1142.
Antunes, P., Reu, C., Sousa, J.C, Pestana, N., Peixe, L. (2002). Incidence and susceptibility to antimicrobial agents of Listeria spp. and Listeria monocytogenes isolated from poultry carcasses in Porto, Portugal. Journal of Food Protection 65(12), 1888-1893.
Borucki,M., and Call, D.R. (2003).Listeria monocytogenes serotype identification using PCR. Journal of Clinical Microbiology 41, 5537–5540.
Cabrita, P., Fonseca, C., Freitas, R., Carreira, R., Capelo, J.L., Trigo, M.J., Ferreira, R.B., Brito, L. (2010). Asecretomebased methodology may provide a better characterization of the virulence of Listeria monocytogenes: Preliminary results. Talanta 83, 457–463.
Doijad, S., Barbuddhe, S.B., Garg, S., Kalekar, S., Rodrigues, J., D’Costa, D., Bhosle, S., Chakraborty, T. (2011). Incidence and genetic variability of Listeria species from three milk processing plants. Food Control 22, 1900-1904.
Doumith, M., Buchrieser, C., Glaser, P., Jaquet, C., Martin, P. (2004). Differentiation of the major Listeria monocytogenesserovars by Multiplex PCR. Journal of Clinical Microbiology 42, 3819-3822.
Engelbrecht, F., Chun, S.K., Ochs, C., Hess, J., Lottspeich, F., Goebel, W., and Sokolovic, Z. (1996). A new prfAregulated gene of Listeria monocytogenes encoding a small, secreted protein which belongs to the family of internalins. Molecular Microbiology 21, 823-837.
Fallah, A.A., Saei-Dehkordi, S.S., Rahnama, M., Tahmasby, H., Mahzounieh, M.R. (2012). Prevalence and antimicrobial resistance patterns of Listeria species isolated from poultry products marketed in Iran. Food Control 28, 327-332.
Faverom, S., Mcdade, J. J., Robertsen, J. A., Hoffman, R. K., & Edwards, R. W. (1968). Microbiological sampling of surfaces. Journal of applied Bacteriology 31, 336-343.
Gelbicova, T., & Karpiskova, R. (2012). Outdoor environment as a source of Listeriamonocytogenes in food chain. Czech Journal of Food Sciences 30, 83-88.
Gouin, E., Mengaud, J., and Cossart, P. (1994). The virulence gene cluster of Listeria monocytogenes is also present in Listeria ivanovii, an animal pathogen, and Listeriaseeligeri, a nonpathogenic species. Infection and Immunity 62, 3550-3553.
Goulet, V. (2006). Surveillance of human listeriosis in France, 2001-2003. Eurosurveillance 11, 79-81.
Guerra, M.M., McLauchin, J., & Bernardo, F.A. (2001). Listeria in ready-to-eat and unprocessed foods produced in Portugal. Food Microbiology 18, 423-429.
Hitchins A.D. & Jinneman, K. (2013). Detection and Enumeration of Listeria monocytogenes in Foods. In Bacteriological Analytical Manual. Available online at: http://www.fda.gov/food/foodscienceresea rch/laboratorymethods/ucm071400.htm
Jacquet, C., Gouin, E., Jeannel, D., Cossart, P., Rocourt, J. (2002). Expression of ActA, Ami, InlB, and listeriolysin O in Listeria monocytogenes of human and food origin. Applied and Environmental Microbiology 68, 616–622.
Jamali, H., Radmehr, B., & Thong, K. L. (2013). Prevalence, characterisation, andantimicrobial resistance of Listeria species and Listeria monocytogenes isolates from raw milk in farm bulk tanks. Food Control 34, 121-125.
Jamali, H., and Thong, K.L. (2014). Genotypic characterization and antimicrobial resistance of Listeria monocytogenes from ready-to-eat foods. Food Control 44, 1-6.
Liu, D. (2006). Identification, subtyping and virulence determination of Listeriamonocytogenes, in important foodborne pathogen. Journal of Medical Microbiology 55, 645-659.
Liu, D., Ainsworth, A. J., Austin, F.W., & Lawrence, M. L. (2004a). Use of PCR primers derived from a putative transcriptional regulator gene for species-specific identification of Listeria monocytogenes. International Journal of Food Microbiology 91, 297–304.
Liu, D., Ainsworth, A.J., Austin, F.W., Lawrence, M.L. (2003a). Characterization of virulent and avirulent Listeria monocytogenes strains by PCR amplification of putative transcriptional regulator and internalin genes. Journal of Medical Microbiology 52, 1065–1070.
Liu, D., Lawrence, M.L., Austin, F.W., Ainsworth, A.J. (2007). A multiplex PCR for species and virulence-specific determination of Listeria monocytogenes. Journal of Microbiological Methods 71, 133–140.
Mammina, C., Aleo, A., Romani, C., Pellissier, N., Nicoletti, P., Pecile, P., et al. (2009). Characterization of Listeria monocytogenes isolates from human listeriosis cases in Italy. Journal of Clinical Microbiology 47, 2925-2930.
Martins, E. A., & Leal Germano, P. M. (2011). Listeria monocytogenes in ready-to-eat, sliced, cooked ham and salami products, marketed in the city of São Paulo, Brazil: occurrence, quantification, and serotyping. Food Control 22, 297-302.
Miettinen, M,K., Palmu, L., Björkroth, K.J., & Korkeala, H. (2001). Prevalence of Listeriamonocytogenes in broilers at the abattoir, processing plant, and retail level. Journal of Food Protection 64, 994-999.
Pagadala, S., Parveen, S., Rippen, T., Luchansky, J.B., Call, J.E., Tamplin, M.L., Porto-Fett, ACS , (2012). Prevalence, characterization and sources of Listeria monocytogenes in blue crab (Callinectussapidus) meat and blue crab processing plants. FoodMicrobiology31,263-270.
Praakle-Amin, A., Hänninen, M.L., & Korkeala, H. (2006). Prevalence and genetic characterization of Listeria monocytogenes in retail broiler meat in Estonia. Journal of Food Protection 69, 436-440.
Rossi, .M.L., Paiva, A., Tornese, M., Chianelli, S., Troncoso, A. (2008). Listeriamonocytogenes outbreaks: a review of the routes that favor bacterial presence. Revistachilena de infectologia 25(5), 328-335.
Sabet, C., Lecuit, M., Cabanes, D., Cossart, P., Bierne, H. (2005). LPXTG protein InlJ, a newly identified internalin involved in Listeria monocytogenes virulence. Infection and Immunity 73, 6912–6922.
Sakaridis, I., Soultos, N., Iossifidou, E., Papa, A., Ambrosiadis, I., & Koidis, P. (2011). Prevalence and antimicrobial resistance of Listeria monocytogenes isolated in chicken slaughterhouses in northern Greece. Journal of Food Protection 74, 1017-1021.
Sant'Ana, A.S., Igarashi, M.C., Landgraf, M., Destro, M.T., Franco, BDGM. (2012). Prevalence, populations and pheno- and genotypic characteristics of Listeria monocytogenes isolated from ready-toeat vegetables marketed in São Paulo, Brazil. International Journal of Food Microbiology155, 1–9.
Vazquez-Boland, J.A., Kuhn, M., Berche, P., Chakraborty, T., Dominguez-Bernal, G., Goebel, W., Gonzalez-Zorn, B., Wehland, J., Kreft, J. (2001). Listeria pathogenesis and molecular virulence determinants. Clinical Microbiology Reviews 14, 584-640.
Wernar, K., Heuvelman, K., Notermans, S., Domann, E., Leimeister-Wachter, M., & Chakraborty, T. (1992). Suitability of the prfA gene, which encodes a regulator of virulence genes in Listeria monocytogenes, in the identification of pathogenic Listeria spp. Applied and Environmental Microbiology 58, 765– 768.
Wiedmann, M., Bruce, J.L., Knorr, R., Bodis, M., Cole, E.M., McDowell, C.I., McDonough, P.L., and Batt, C.A. (1996). Ribotype diversity of Listeria monocytogenes strains associated with outbreaks of listeriosis in ruminants. Journal of Clinical Microbiology 34, 1086–1090.
Zhang, Y., Yeh, E., Hall, G., Cripe, J., Bhagwat, A.A., & Meng, J. (2007). Characterization of Listeria monocytogenes isolated from retail foods. International Journal of Food Microbiology 113, 47-53.